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B-CPAP\uff08\u8001\u5e74\u4e73\u5934\u72b6\uff09\uff0c\u4f53\u73b0\u6210\u5e74\u5973\u6027\u4e73\u5934\u72b6\u7532\u72b6\u817a\u764c\u7684\u5178\u578b\u5f62\u6001\uff1b\u4e0e TT\uff08\u677e\u6563\u4e0a\u76ae\u6837\u3001\u542b\u5206\u6ccc\u9897\u7c92\uff09\u76f8\u6bd4\uff0c\u6392\u5217\u66f4\u7d27\u5bc6\u4e14\u65e0\u795e\u7ecf\u5185\u5206\u6ccc\u7279\u5f81\uff0c\u4e0e B-CPAP\uff08\u6df7\u5408\u5f62\u6001\uff09\u76f8\u6bd4\uff0c\u5f62\u6001\u66f4\u5747\u4e00\u4e14\u5177\u5907\u5e74\u8f7b\u80bf\u7624\u7ec6\u80de\u7684\u6d3b\u529b\u7279\u5f81\uff1b\u53ef\u901a\u8fc7 \u201c\u4e0a\u76ae\u6837 + \u4e73\u5934\u72b6\u764c\u6807\u5fd7\u7269\uff08CK19\u3001TTF-1\u3001Tg\uff09\u9633\u6027\u201d \u5feb\u901f\u9274\u522b\uff0c\u540c\u65f6\u4e0e TT \u901a\u8fc7\u75c5\u7406\u4e9a\u578b\u6807\u5fd7\u7269\uff08\u5982 CK19 vs \u964d\u8840\u9499\u7d20\uff09\u3001\u4e0e B-CPAP \u901a\u8fc7\u5e74\u9f84\u76f8\u5173\u6807\u5fd7\u7269\uff08\u5982 p16 \u8868\u8fbe\u5dee\u5f02\uff09\u8fdb\u4e00\u6b65\u533a\u5206\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7ec6\u80de\u4ee3\u6570\uff1a10 \u4ee3\u4ee5\u5185\uff08\u4f4e\u4ee3\u6570\u7ec6\u80de\u80fd\u66f4\u597d\u4fdd\u7559 \u201c\u4e73\u5934\u72b6\u7279\u6027\u3001\u4e0a\u76ae\u5f62\u6001\u3001\u6210\u5e74\u80bf\u7624\u5c5e\u6027\u201d \u6838\u5fc3\u5c5e\u6027\uff0c\u5efa\u8bae\u4f20\u4ee3\u6b21\u6570\u4e25\u683c\u63a7\u5236\u5728 10 \u4ee3\u4ee5\u5185\uff0c\u9632\u6b62\u957f\u671f\u4f20\u4ee3\u5bfc\u81f4\u7279\u6027\u6f02\u79fb\uff0c\u5982\u4e0a\u76ae\u5f62\u6001\u5411\u68ad\u5f62\u8f6c\u53d8\u3001\u4e73\u5934\u72b6\u6807\u5fd7\u7269\uff08CK19\u3001TTF-1\uff09\u8868\u8fbe\u4e0b\u964d\u3001\u589e\u6b96\u901f\u5ea6\u5f02\u5e38\uff08\uff1c25 \u5c0f\u65f6\u6216\uff1e40 \u5c0f\u65f6\uff09\uff0c\u786e\u4fdd\u7ec6\u80de\u59cb\u7ec8\u8d34\u5408\u6210\u5e74\u5973\u6027\u4e73\u5934\u72b6\u7532\u72b6\u817a\u764c\u7684\u5b9e\u9a8c\u9700\u6c42\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u80cc\u666f\u7b80\u4ecb\uff1aTPC-1 \u7ec6\u80de\u4e3a\u6210\u5e74\u5973\u6027\u7532\u72b6\u817a\u4e73\u5934\u72b6\u764c\u6765\u6e90\u7ec6\u80de\u7cfb\uff0c\u6838\u5fc3\u7279\u6027\u53ca\u5e94\u7528\u5982\u4e0b\uff1a\u2460\u5e74\u9f84\u4e0e\u4e9a\u578b\u7279\u8272\uff1a\u4f5c\u4e3a\u6210\u5e74\u5973\u6027\u4e73\u5934\u72b6\u7532\u72b6\u817a\u764c\u4ee3\u8868\u6a21\u578b\uff0c\u586b\u8865 \u201c\u6210\u5e74 vs \u8001\u5e74\u201d \u4e73\u5934\u72b6\u764c\u7814\u7a76\u7a7a\u767d\uff0c\u53ef\u7528\u4e8e\u63a2\u7d22\u5e74\u9f84\u76f8\u5173\u7684\u80bf\u7624\u8fdb\u5c55\u5dee\u5f02\uff08\u5982\u6210\u5e74\u80bf\u7624\u7684\u4f4e\u4fb5\u88ad\u6027 vs \u8001\u5e74\u80bf\u7624\u7684\u9ad8\u6076\u6027\uff09\uff1b\u2461\u5e94\u7528\u573a\u666f\uff1a\u5e7f\u6cdb\u7528\u4e8e\u7532\u72b6\u817a\u4e73\u5934\u72b6\u764c\u57fa\u7840\u751f\u7269\u5b66\u7814\u7a76\uff08\u5982\u7ec6\u80de\u5468\u671f\u8c03\u63a7\u3001\u4fe1\u53f7\u901a\u8def\u5206\u6790\uff09\u3001\u7532\u72b6\u817a\u764c\u4e9a\u578b\u5bf9\u6bd4\u5b9e\u9a8c\uff08\u4e0e TT \u9ad3\u6837\u764c\uff09\u3001\u6210\u5e74\u5973\u6027\u7532\u72b6\u817a\u764c\u836f\u7269\u7b5b\u9009\uff08\u5982\u9776\u5411 RET \u878d\u5408\u86cb\u767d\u7684\u836f\u7269\uff09\uff0c\u540c\u65f6\u56e0\u5e38\u89c4\u57f9\u517b\u6761\u4ef6\uff0c\u9002\u5408\u5f00\u5c55\u4e34\u5e8a\u6837\u672c\u76f8\u5173\u6027\u5b9e\u9a8c\uff08\u5982\u4e0e\u6210\u5e74\u60a3\u8005\u7ec4\u7ec7\u6837\u672c\u7684\u57fa\u56e0\u8868\u8fbe\u5bf9\u6bd4\uff09\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">STR \u9274\u5b9a\uff1aSTR \u9274\u5b9a\u6b63\u786e\uff08\u6838\u5fc3\u4fdd\u969c\uff1a\u2460\u901a\u8fc7\u81f3\u5c11 16 \u4e2a\u6838\u5fc3 STR \u4f4d\u70b9\u68c0\u6d4b\uff08\u5982 Amelogenin\u3001CSF1PO\u3001D13S317 \u7b49\uff09\uff0c\u7ed3\u679c\u4e0e\u7532\u72b6\u817a\u4e73\u5934\u72b6\u764c\u7ec6\u80de\u6807\u51c6\u56fe\u8c31\u4e00\u81f4\uff0c\u65e0 TT\u3001B-CPAP \u7684 STR \u4f4d\u70b9\u7279\u5f81\uff0c\u6392\u9664\u4ea4\u53c9\u6c61\u67d3\uff1b\u2461\u53ef\u901a\u8fc7\u5bf9\u6bd4\u56fd\u5185\u7532\u72b6\u817a\u7ec6\u80de\u5e93\uff08\u5982\u4e2d\u56fd\u533b\u5b66\u79d1\u5b66\u9662\u57fa\u7840\u533b\u5b66\u7814\u7a76\u6240\u7ec6\u80de\u8d44\u6e90\u4e2d\u5fc3\uff09\u7684\u6210\u5e74\u5973\u6027\u4e73\u5934\u72b6\u7532\u72b6\u817a\u764c\u7ec6\u80de STR \u6570\u636e\uff0c\u8fdb\u4e00\u6b65\u9a8c\u8bc1\u8eab\u4efd\u53ca\u5e74\u9f84\u76f8\u5173\u7279\u5f81\uff0c\u4e0e TT \u7684 \u201c\u53cc\u533a\u95f4\u6838\u578b STR\u201d\u3001B-CPAP \u7684 \u201c\u8001\u5e74\u4e73\u5934\u72b6 STR\u201d \u5f62\u6210\u5dee\u5f02\uff0c\u786e\u4fdd\u5b9e\u9a8c\u7528\u7ec6\u80de\u8eab\u4efd\u7cbe\u51c6\u4e14\u5e74\u9f84\u3001\u4e9a\u578b\u7279\u6027\u660e\u786e\uff0c\u907f\u514d\u56e0\u4e9a\u578b\u6df7\u6dc6\u6216\u5e74\u9f84\u8bef\u5224\u5bfc\u81f4\u5b9e\u9a8c\u7ed3\u8bba\u504f\u5dee\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u751f\u7269\u5b89\u5168\u7b49\u7ea7\uff1a1\uff08\u4e0e TT\u3001B-CPAP \u4e00\u81f4\uff0c\u4f4e\u4e8e BEAS-2B \u7684 2 \u7ea7\uff1b\u64cd\u4f5c\u65f6\u9700\u9075\u5faa\u751f\u7269\u5b89\u5168\u4e00\u7ea7\u5b9e\u9a8c\u5ba4\u9632\u62a4\u89c4\u8303\uff0c\u5b9e\u9a8c\u5e9f\u5f03\u7269\u9700\u7ecf 121\u2103\u9ad8\u538b\u706d\u83cc 30 \u5206\u949f\u5904\u7406\uff0c\u907f\u514d\u7ec6\u80de\u6b8b\u7559\u6c61\u67d3\uff1b\u82e5\u6d89\u53ca\u4e34\u5e8a\u6837\u672c\u5173\u8054\u5b9e\u9a8c\uff0c\u9700\u4e25\u683c\u9075\u5b88\u751f\u7269\u6837\u672c\u4f26\u7406\u89c4\u8303\uff0c\u9632\u6b62\u6837\u672c\u4ea4\u53c9\u6c61\u67d3\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7ec6\u80de\u89c4\u683c\uff1a1\u00d710\u2076cells\/T25 \u57f9\u517b\u74f6\u6216\u8005 1mL \u51bb\u5b58\u7ba1\u5305\u88c5\uff08\u8ba1\u6570\u65f6\u56e0\u4e0a\u76ae\u7ec6\u80de\u6392\u5217\u89c4\u6574\uff0c\u8f7b\u67d4\u5439\u6253\u5373\u53ef\u83b7\u5f97\u5355\u7ec6\u80de\u60ac\u6db2\uff0c\u8ba1\u6570\u51c6\u786e\u6027\u9ad8\uff08\u8bef\u5dee\uff1c5%\uff09\uff0c\u65e0 TT \u7684\u5206\u6ccc\u9897\u7c92\u5e72\u6270\u3001B-CPAP \u7684\u5f62\u6001\u5f02\u8d28\u6027\u8ba1\u6570\u504f\u5dee\u95ee\u9898\uff1b\u51bb\u5b58\u7ba1\u542b 10% \u4e8c\u7532\u57fa\u4e9a\u781c\uff08DMSO\uff09\u65e0\u8840\u6e05\u51bb\u5b58\u6db2\uff0c\u590d\u82cf\u540e 48 \u5c0f\u65f6\u5b58\u6d3b\u7387\u226585%\uff0c\u4e14\u4e0a\u76ae\u6837\u5f62\u6001\u3001\u4e73\u5934\u72b6\u7279\u6027\u65e0\u663e\u8457\u6539\u53d8\uff1bT25 \u74f6\u53d1\u8d27\u65f6\u7ec6\u80de\u5bc6\u5ea6\u8fbe 70%-75%\uff0c\u6807\u6ce8 \u201c\u6210\u5e74\u5973\u6027\u4e73\u5934\u72b6\u7532\u72b6\u817a\u764c\uff0cSTR \u9274\u5b9a\u6b63\u786e\uff0c\u5efa\u8bae\u68c0\u6d4b CK19\/TTF-1\u201d\uff0c\u63d0\u9192\u7528\u6237\u5229\u7528\u6210\u5e74\u5e74\u9f84\u53ca\u4e73\u5934\u72b6\u4e9a\u578b\u7279\u6027\u5f00\u5c55\u5b9e\u9a8c\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u652f\u539f\u4f53\u68c0\u6d4b\uff1a\u65e0\uff08\u6bcf\u6279\u6b21\u7ec6\u80de\u901a\u8fc7 PCR \u6cd5\u68c0\u6d4b\u652f\u539f\u4f53\uff08\u8986\u76d6\u80ba\u708e\u652f\u539f\u4f53\u3001\u4eba\u578b\u652f\u539f\u4f53\u3001\u89e3\u8132\u652f\u539f\u4f53\uff09\uff0c\u7ed3\u679c\u4e3a\u9634\u6027\uff0c\u907f\u514d\u652f\u539f\u4f53\u6c61\u67d3\u5f71\u54cd\u7ec6\u80de\u589e\u6b96\u3001\u5f62\u6001\u53ca\u540e\u7eed\u5b9e\u9a8c\uff08\u5982\u836f\u7269\u7b5b\u9009\u3001\u8f6c\u67d3\uff09\uff0c\u4fdd\u969c\u4e0e TT\u3001B-CPAP \u5e73\u884c\u5b9e\u9a8c\u7684\u53ef\u9760\u6027\uff0c\u5c24\u5176\u9002\u5408\u591a\u7ec6\u80de\u7cfb\u540c\u6b65\u5f00\u5c55\u7684\u4e9a\u578b\u5bf9\u6bd4\u5b9e\u9a8c\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u4fdd\u85cf\u673a\u6784\uff1a\u672a\u660e\u786e\u6807\u6ce8\uff08\u5efa\u8bae\u5b9e\u9a8c\u524d\u54a8\u8be2\u4f9b\u5e94\u5546\u83b7\u53d6\u4fdd\u85cf\u673a\u6784\u4fe1\u606f\uff08\u5982 ATCC\u3001DSMZ \u6216\u56fd\u5185\u7532\u72b6\u817a\u7ec6\u80de\u5e93\uff09\uff0c\u901a\u8fc7\u4fdd\u85cf\u673a\u6784\u6570\u636e\u5e93\u67e5\u8be2\u7ec6\u80de\u8be6\u7ec6\u80cc\u666f\uff08\u5982\u5177\u4f53\u5e74\u9f84\u3001\u662f\u5426\u4e3a\u539f\u53d1\u7076\u3001\u662f\u5426\u542b RET \u878d\u5408\u7b49\u4e73\u5934\u72b6\u764c\u7279\u5f81\u7a81\u53d8\uff09\uff0c\u8865\u5145\u5b8c\u5584\u7ec6\u80de\u57fa\u7840\u4fe1\u606f\uff0c\u63d0\u5347\u5b9e\u9a8c\u53ef\u8ffd\u6eaf\u6027\u4e0e\u7ed3\u679c\u91cd\u590d\u6027\uff0c\u4e0e TT \u7684\u56fd\u5185\u6743\u5a01\u4fdd\u85cf\u3001B-CPAP \u7684 DSMZ \u4fdd\u85cf\u5f62\u6210\u4e92\u8865\uff0c\u4fbf\u4e8e\u56fd\u5185\u5916\u79d1\u7814\u534f\u4f5c\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u57f9\u517b\u57fa\uff1a90% DMEM+10% FBS+PS\uff08\u9752\u9709\u7d20 &#8211; \u94fe\u9709\u7d20\uff09\uff08\u6838\u5fc3\u7279\u8272\uff1a\u2460\u91c7\u7528 DMEM \u57f9\u517b\u57fa\uff0c\u4e0e TT \u7684 F-12K\u3001B-CPAP \u7684 RPMI-1640 \u5f62\u6210\u663e\u8457\u5dee\u5f02\uff0c\u542b\u9ad8\u7cd6\uff084.5g\/L\uff09\u53ca\u9002\u914d\u6210\u5e74\u4e73\u5934\u72b6\u7532\u72b6\u817a\u764c\u7ec6\u80de\u7684\u8425\u517b\u914d\u6bd4\uff0c\u652f\u6301\u5176\u4e0a\u76ae\u5f62\u6001\u7ef4\u6301\u53ca\u4e73\u5934\u72b6\u764c\u76f8\u5173\u86cb\u767d\uff08\u5982 CK19\u3001TTF-1\uff09\u5408\u6210\uff1b\u246110% FBS \u9700\u9009\u62e9\u4f4e\u5185\u6bd2\u7d20\u3001\u6279\u6b21\u95f4\u5dee\u5f02\u5c0f\u7684\u4f18\u8d28\u80ce\u725b\u8840\u6e05\uff0c\u907f\u514d\u5f71\u54cd\u7ec6\u80de\u589e\u6b96\u5747\u4e00\u6027\u53ca\u4e73\u5934\u72b6\u7279\u6027\uff0c\u65e0\u9700\u50cf TT \u90a3\u6837\u8003\u8651\u5206\u6ccc\u529f\u80fd\u76f8\u5173\u8425\u517b\u9700\u6c42\uff1b\u2462PS \u7ec8\u6d53\u5ea6\u4e3a 100U\/mL \u9752\u9709\u7d20\u3001100\u03bcg\/mL \u94fe\u9709\u7d20\uff0c\u6709\u6548\u9884\u9632\u7ec6\u83cc\u6c61\u67d3\uff0c\u4e0d\u5f71\u54cd\u7532\u72b6\u817a\u4e73\u5934\u72b6\u764c\u8868\u578b\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u57f9\u517b\u6761\u4ef6\uff1a\u6c14\u76f8\uff1a95% \u7a7a\u6c14 + 5% \u4e8c\u6c27\u5316\u78b3\uff1b\u6e29\u5ea6\uff1a37\u2103\uff08\u4e0e TT\u3001B-CPAP \u7684\u5e38\u89c4 CO\u2082\u57f9\u517b\u6761\u4ef6\u4e00\u81f4\uff0c\u6e7f\u5ea6\u7ef4\u6301\u5728 70%-80%\uff0c\u5b9a\u671f\u66f4\u6362\u6c34\u76d8\uff0c\u9632\u6b62\u57f9\u517b\u57fa\u84b8\u53d1\u5bfc\u81f4\u6e17\u900f\u538b\u5347\u9ad8\uff1b\u57f9\u517b\u7bb1\u5185\u907f\u514d\u5267\u70c8\u6643\u52a8\uff0c\u9632\u6b62\u8d34\u58c1\u7ec6\u80de\u8131\u843d\uff1b\u56e0\u4f7f\u7528 DMEM \u57f9\u517b\u57fa\uff0c\u4e0e\u5176\u4ed6\u7532\u72b6\u817a\u764c\u7ec6\u80de\uff08TT\/B-CPAP\uff09\u4ec5\u9700\u6ce8\u610f\u57f9\u517b\u57fa\u5dee\u5f02\uff0c\u57f9\u517b\u73af\u5883\u517c\u5bb9\u6027\u9ad8\uff0c\u4fbf\u4e8e\u591a\u7ec6\u80de\u682a\u5e76\u884c\u5f00\u5c55 \u201c\u4e9a\u578b \/ \u5e74\u9f84\u5bf9\u6bd4\u201d \u5b9e\u9a8c\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u51bb\u5b58\u6761\u4ef6\uff1a\u65e0\u8840\u6e05\u51bb\u5b58\u6db2\uff0c\u6db2\u6c2e\u50a8\u5b58\uff08\u51bb\u5b58\u64cd\u4f5c\u9002\u914d\u4e0a\u76ae\u7ec6\u80de\u6837\u5f62\u6001\u53ca\u6210\u5e74\u80bf\u7624\u7279\u6027\uff1a\u2460\u9009\u62e9\u5bf9\u6570\u751f\u957f\u671f\u4e2d\u671f\u7ec6\u80de\uff08\u5bc6\u5ea6 75%\uff0c\u6d3b\u6027\u6700\u4f73\u3001\u4e73\u5934\u72b6\u7279\u6027\u7a33\u5b9a\uff09\uff0c\u7528 0.25% \u80f0\u9176 &#8211; EDTA \u5b75\u80b2 2-3 \u5206\u949f\uff0c\u955c\u4e0b\u89c1\u7ec6\u80de\u95f4\u9699\u589e\u5927\u5373\u53ef\u7ec8\u6b62\uff0c\u8f7b\u67d4\u5439\u6253\u907f\u514d\u7ec6\u80de\u7834\u788e\uff1b\u24611000rpm \u79bb\u5fc3 5 \u5206\u949f\u6536\u96c6\u7ec6\u80de\uff0c\u7528\u9884\u51b7\u7684\u65e0\u8840\u6e05\u51bb\u5b58\u6db2\u91cd\u60ac\uff0c\u8c03\u6574\u6d53\u5ea6\u81f3 2.5\u00d710\u2076cells\/mL\uff08\u4e0e TT\u3001B-CPAP \u4e00\u81f4\uff09\uff1b\u2462\u51bb\u5b58\u6d41\u7a0b\uff1a4\u2103\u5e73\u8861 30 \u5206\u949f\u2192-20\u2103\u653e\u7f6e 2 \u5c0f\u65f6\uff08\u7a0b\u5e8f\u964d\u6e29\u76d2\uff09\u2192-80\u2103\u8fc7\u591c\u2192\u6db2\u6c2e\u4fdd\u5b58\uff1b\u590d\u82cf\u65f6 37\u2103\u6c34\u6d74 1 \u5206\u949f\u878d\u5316\uff0c\u79bb\u5fc3\u540e\u7528\u9884\u70ed\u7684 DMEM \u57f9\u517b\u57fa\u91cd\u60ac\uff0c\u76f4\u63a5\u63a5\u79cd\uff0c\u907f\u514d DMSO \u5f71\u54cd\u4e73\u5934\u72b6\u764c\u76f8\u5173\u86cb\u767d\uff08\u5982 Tg\uff09\u8868\u8fbe\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7ec6\u80de\u8d27\u671f\uff1a\u73b0\u8d27\uff0c1 \u5468\u5de6\u53f3\uff08\u5e93\u5b58\u7ec6\u80de\u5747\u7ecf\u8fc7 DMEM \u57f9\u517b\u57fa\u9002\u5e94\u6027\u57f9\u517b\u3001\u652f\u539f\u4f53\u68c0\u6d4b\u53ca\u7ec6\u80de\u5f62\u6001\u9a8c\u8bc1\uff08\u4e0a\u76ae\u6837\uff09\uff0c\u6838\u5fc3\u57f9\u517b\u7279\u6027\u7a33\u5b9a\uff0c\u4e0e TT\u3001B-CPAP \u7684 1 \u5468\u8d27\u671f\u4e00\u81f4\uff1b\u4e0b\u5355\u540e 48 \u5c0f\u65f6\u5185\u53ef\u5b89\u6392\u53d1\u8d27\uff0c\u53ef\u4e0e TT\uff08\u9ad3\u6837\uff09\u3001B-CPAP\uff08\u8001\u5e74\u4e73\u5934\u72b6\uff09\u540c\u6b65\u4e0b\u5355\uff0c\u4fdd\u969c \u201c\u7532\u72b6\u817a\u764c\u4e9a\u578b \/ \u5e74\u9f84\u68af\u5ea6\u201d \u5e73\u884c\u5b9e\u9a8c\u7684\u65f6\u95f4\u4e00\u81f4\u6027\uff0c\u65e0\u9700\u989d\u5916\u7b49\u5f85\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u8fd0\u8f93\u65b9\u5f0f\uff1a\u590d\u82cf\u53d1\u8d27\uff08T25 \u74f6\u514d\u8fd0\u8f93\u8d39\u7528\uff0c\u91c7\u7528\u9632\u9707\u6052\u6e29\u6ce1\u6cab\u7bb1\uff0c\u5185\u7f6e 2-8\u2103\u51b0\u888b\u53ca\u6e29\u5ea6\u76d1\u6d4b\u5361\uff0c\u8fd0\u8f93\u8fc7\u7a0b\u4e2d\u7528\u56fa\u5b9a\u652f\u67b6\u9632\u6b62\u57f9\u517b\u74f6\u6643\u52a8\uff0c\u51cf\u5c11\u4e0a\u76ae\u6837\u8d34\u58c1\u7ec6\u80de\u8131\u843d\uff1b\u5230\u8fbe\u540e\u7acb\u5373\u653e\u5165 5% CO\u2082\u57f9\u517b\u7bb1\uff0c24 \u5c0f\u65f6\u540e\u89c2\u5bdf\u7ec6\u80de\u8d34\u58c1\u72b6\u6001\uff0c48 \u5c0f\u65f6\u5185\u9996\u6b21\u6362\u6db2\uff08\u4f7f\u7528 90% DMEM+10% FBS+PS \u57f9\u517b\u57fa\uff09\uff0c\u786e\u4fdd\u7ec6\u80de\u9002\u5e94\u73af\u5883\uff0c\u907f\u514d\u8fd0\u8f93\u5e94\u6fc0\u5f71\u54cd\u6210\u5e74\u80bf\u7624\u7ec6\u80de\u7684\u6d3b\u529b\uff09\/ \u51bb\u5b58\u53d1\u8d27\uff08\u9700\u52a0\u5e72\u51b0\u8fd0\u8f93\u8d39\u7528\uff0c\u5e72\u51b0\u7528\u91cf\u6309\u8fd0\u8f93\u65f6\u957f\u8ba1\u7b97\uff0c\u56fd\u5185\u8fd0\u8f93\u4e00\u822c\u914d\u5907 1kg \u5e72\u51b0\uff0c\u786e\u4fdd\u5230\u8fbe\u65f6\u5e72\u51b0\u5269\u4f59\u91cf\u226560%\uff0c\u7ef4\u6301\u7ec6\u80de\u51bb\u5b58\u72b6\u6001\uff0c\u4fdd\u969c\u4e73\u5934\u72b6\u7279\u6027\u65e0\u6539\u53d8\uff09 \u987a\u4e30\u5feb\u9012\uff08\u4f18\u5148\u9009\u62e9\u6b21\u6668\u8fbe\u670d\u52a1\uff0c\u6700\u5927\u9650\u5ea6\u7f29\u77ed\u8fd0\u8f93\u65f6\u95f4\uff0c\u964d\u4f4e\u7ec6\u80de\u6d3b\u6027\u635f\u5931\u53ca\u4e0a\u76ae\u5f62\u6001\u6539\u53d8\u98ce\u9669\uff0c\u5c24\u5176\u9002\u5408\u5bf9\u8fd0\u8f93\u6761\u4ef6\u654f\u611f\u7684\u6210\u5e74\u80bf\u7624\u7ec6\u80de\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u4f9b\u5e94\u9650\u5236\uff1a\u4ec5\u9650\u4e8e\u79d1\u5b66\u7814\u7a76\uff0c\u7edd\u4e0d\u53ef\u4f5c\u4e3a\u52a8\u7269\u6216\u4eba\u7c7b\u75be\u75c5\u7684\u6cbb\u7597\u4ea7\u54c1\u4f7f\u7528\uff08\u4e0e TT\u3001B-CPAP \u7b49\u7ec6\u80de\u9650\u5236\u6761\u6b3e\u4e00\u81f4\uff0c\u4e25\u7981\u7528\u4e8e\u4e34\u5e8a\u6cbb\u7597\u3001\u7ec6\u80de\u6cbb\u7597\u3001\u7f8e\u5bb9\u53ca\u5176\u4ed6\u975e\u79d1\u7814\u7528\u9014\uff1b\u5b9e\u9a8c\u8fc7\u7a0b\u4e2d\u9700\u59a5\u5584\u5904\u7406\u542b\u7532\u72b6\u817a\u764c\u7ec6\u80de\u7684\u57f9\u517b\u57fa\uff0c\u907f\u514d\u73af\u5883\u66b4\u9732\uff0c\u82e5\u540e\u7eed\u8865\u5145\u7ec6\u80de\u7a81\u53d8\u4fe1\u606f\uff08\u5982 RET \u878d\u5408\uff09\uff0c\u9700\u5728\u5b9e\u9a8c\u8bb0\u5f55\u4e2d\u660e\u786e\u6807\u6ce8\uff0c\u786e\u4fdd\u6570\u636e\u5b8c\u6574\u6027\u53ca\u5b9e\u9a8c\u53ef\u590d\u73b0\u6027\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7279\u522b\u8bf4\u660e\uff1a\u4ee5\u4e0b\u7ec6\u80de\u57f9\u517b\u51bb\u5b58\u5904\u7406\u4ec5\u4f9b\u53c2\u8003\uff0c\u5177\u4f53\u64cd\u4f5c\u6b65\u9aa4\u4ee5\u968f\u8d27\u4ea7\u54c1\u8bf4\u660e\u4e66\u4e3a\u4e3b\uff1b\u8be5\u7ec6\u80de\u5b9e\u9a8c\u64cd\u4f5c\u9700\u91cd\u70b9\u5173\u6ce8\uff1a\u2460\u4e73\u5934\u72b6\u7279\u6027\u9a8c\u8bc1\uff1a\u9996\u6b21\u57f9\u517b\u65f6\uff0c\u5efa\u8bae\u901a\u8fc7\u514d\u75ab\u8367\u5149\u6216 Western Blot \u68c0\u6d4b\u4e73\u5934\u72b6\u7532\u72b6\u817a\u764c\u7279\u5f02\u6027\u6807\u5fd7\u7269\uff0c\u5982\uff1aa. \u6838\u5fc3\u6807\u5fd7\u7269\uff08CK19\u3001TTF-1\u3001Tg\uff09\uff0c\u9884\u8ba1\u5747\u5448\u9633\u6027\u8868\u8fbe\uff1bb. \u7a81\u53d8\u76f8\u5173\u6807\u5fd7\u7269\uff08\u5982 RET \u878d\u5408\u86cb\u767d\uff0c\u5e38\u89c1\u4e8e\u4e73\u5934\u72b6\u764c\uff09\uff0c\u53ef\u901a\u8fc7 PCR \u6216\u6d4b\u5e8f\u9a8c\u8bc1\uff0c\u660e\u786e\u7ec6\u80de\u662f\u5426\u542b\u4e73\u5934\u72b6\u764c\u5178\u578b\u7a81\u53d8\uff0c\u63d0\u5347\u5b9e\u9a8c\u7cbe\u51c6\u6027\uff1b\u53ef\u8bbe\u7f6e B-CPAP\uff08\u8001\u5e74\u4e73\u5934\u72b6\u9633\u6027\u5bf9\u7167\uff09\u3001TT\uff08\u9ad3\u6837\u9634\u6027\u5bf9\u7167\uff09\uff0c\u786e\u4fdd\u68c0\u6d4b\u7279\u5f02\u6027\uff1b\u2461\u5e74\u9f84\u4e0e\u4e9a\u578b\u5bf9\u6bd4\u5b9e\u9a8c\uff1a\u642d\u914d TT\uff08\u8001\u5e74\u9ad3\u6837\uff09\u3001B-CPAP\uff08\u8001\u5e74\u4e73\u5934\u72b6\uff09\u5f00\u5c55\u5b9e\u9a8c\u65f6\uff0c\u53ef\uff1aa. \u68c0\u6d4b\u5e74\u9f84\u76f8\u5173\u57fa\u56e0\uff08\u5982 p16\u3001p21\uff0c\u8001\u5e74\u7ec6\u80de\u8868\u8fbe\u66f4\u9ad8\uff09\uff0c\u5206\u6790\u6210\u5e74\u4e0e\u8001\u5e74\u7532\u72b6\u817a\u764c\u7684\u8870\u8001\u7279\u5f81\u5dee\u5f02\uff1bb. \u5bf9\u6bd4\u75c5\u7406\u4e9a\u578b\u6807\u5fd7\u7269\uff08\u4e73\u5934\u72b6 CK19 vs \u9ad3\u6837\u964d\u8840\u9499\u7d20\uff09\uff0c\u9a8c\u8bc1\u4e9a\u578b\u7279\u5f02\u6027\uff1bc. \u68c0\u6d4b\u589e\u6b96\u76f8\u5173\u57fa\u56e0\uff08\u5982 Ki-67\uff0c\u6210\u5e74\u7ec6\u80de\u589e\u6b96\u6d3b\u6027\u53ef\u80fd\u9ad8\u4e8e\u8001\u5e74\u7ec6\u80de\uff09\uff0c\u63a2\u7d22\u5e74\u9f84\u5bf9\u80bf\u7624\u589e\u6b96\u7684\u5f71\u54cd\uff1b\u2462\u57fa\u7840\u5b9e\u9a8c\u5e94\u7528\uff1a\u56e0\u5e38\u89c4\u57f9\u517b\u6761\u4ef6\u53ca\u4e0a\u76ae\u6837\u5f62\u6001\uff0c\u9002\u5408\u5f00\u5c55\uff1aa. \u8f6c\u67d3\u5b9e\u9a8c\uff1a\u9009\u62e9\u8102\u8d28\u4f53\u8f6c\u67d3\u6cd5\uff08\u5982 Lipofectamine 3000\uff09\uff0c\u8f6c\u67d3\u6548\u7387\u9884\u8ba1 15%-25%\uff08\u53c2\u8003\u540c\u7c7b\u4e73\u5934\u72b6\u764c\u7ec6\u80de\uff09\uff0c\u8f6c\u67d3\u540e 48 \u5c0f\u65f6\u68c0\u6d4b\u76ee\u7684\u57fa\u56e0\u8868\u8fbe\uff0c\u53ef\u7528\u4e8e\u57fa\u56e0\u529f\u80fd\u7814\u7a76\uff1bb. 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